Improving Endoglucanase Activity by Adding the Carbohydrate-Binding Module from Corticium rolfsii
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The carbohydrate-binding module (CBM) is an important domain of most cellulases that playsa key role in the hydrolysis of cellulose. The neutral endoglucanase (EG1) gene wasreconstructed. A redesigned endoglucanase, named EG2, was constructed with a CBMcontaining a linker from Corticium rolfsii (GenBank Accession No. D49448). The redesigned EGgenes were expressed in Escherichia coli, and their characteristics are discussed. Resultsshowed that the degradation of cellulose by EG2 was about double that by EG1. The specificactivities of EG1 and EG2 were tested under optimal conditions, and EG2 had higher activity(169.1 ± 2.74 U/mg) toward CMC-Na than did EG1 (84.0 ± 1.98) in the process of cellulosedegradation. The optimal pH and temperature, pH stability, and heat stability of EG1 and EG2were similar. Results indicated that the CBM plays an essential role in the hydrolysis ofcellulose. We can improve EG’s catalytic power by adding the CBM from Corticium rolfsii.
Abstract
The carbohydrate-binding module (CBM) is an important domain of most cellulases that playsa key role in the hydrolysis of cellulose. The neutral endoglucanase (EG1) gene wasreconstructed. A redesigned endoglucanase, named EG2, was constructed with a CBMcontaining a linker from Corticium rolfsii (GenBank Accession No. D49448). The redesigned EGgenes were expressed in Escherichia coli, and their characteristics are discussed. Resultsshowed that the degradation of cellulose by EG2 was about double that by EG1. The specificactivities of EG1 and EG2 were tested under optimal conditions, and EG2 had higher activity(169.1 ± 2.74 U/mg) toward CMC-Na than did EG1 (84.0 ± 1.98) in the process of cellulosedegradation. The optimal pH and temperature, pH stability, and heat stability of EG1 and EG2were similar. Results indicated that the CBM plays an essential role in the hydrolysis ofcellulose. We can improve EG’s catalytic power by adding the CBM from Corticium rolfsii.
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